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Experimental Protocols for Generation and Evaluation of Articular Cartilage
Published in Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi, Articular Cartilage, 2017
Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi
Common stains and their chemistry used for articular cartilage are described in Section 5.2. The general procedure for preparing frozen sections differs from that for fixed paraffin-embedded sections. However, subsequent staining is similar. Fixation involves chemically cross-linking the components of the cells and tissue to preserve morphology. Fixatives for paraffin embedding include neutral buffered formalin, paraformaldehyde, and Bouin’s fixative. Frozen sections can be produced with or without prior fixation. After embedding the sample and sectioning, unfixed sections should be fixed prior to staining. The use of a microtome or cryotome for sample sectioning is not described here. It should be noted that sectioning requires a significant amount of “art” to produce quality sections.
Immunofluorescence
Published in Guy Cox, Fundamentals of Fluorescence Imaging, 2019
Immunofluorescence can be done on fresh frozen cell preparations (e.g., renal and skin autoimmune diagnosis) or on tissues that have been “fixed.” Fixation preserves cells and tissues in a lifelike condition, inhibits autolytic (self-digestion) and putrefaction changes, and insolubilizes tissue elements for later demonstration. It is important that fixation has minimal effect on antigenic determinates, allowing recognition by appropriate antibodies. Fixatives can be physical or chemical in action. Physical fixatives, for example heating sections or cells on to slides, alcohol, or acetone treatment, preserve tissues by dehydrating cells. Alcohol and acetone possibly also dissolve lipids from cell membranes facilitating antibody access.
Pressure chamber with a viewport and magnet manipulators to study biological samples
Published in Instrumentation Science & Technology, 2018
Maksim Suslov, Alexander Anisimov
The present study reports a new pressure chamber with magnet manipulators to study biological samples. The chamber allows the monitoring of the dynamics of growth rate of plant roots and other biological samples under changes in external pressure. Using the described pressure chamber, the dynamics of maize root growth and photos of the crosscut of root segments under the effect of elevated air pressure were obtained. It becomes possible to treat a sample with chemicals and fix it for microscopy immediately during changes in external pressure to avoid additional side effects, caused, for example, by decompression. The application of the pressure chamber for chemical fixation of the sample for electron microscopy allows microphotos not only of the tissue cross section but cell ultrastructure under pressure. The results of electron microscopy being used, for example, to determine the correlation between cell growth rate and changes in their ultrastructure in respond to external pressure changes or to compare the results of biological experiments with mathematical computer simulation of cell packing in tissues of various biological samples under volume compression. The size of the chamber allows its temperature control within standard thermostats, but also the chamber can potentially be modified, and temperature control can be provided using thermoelectric Peltier moduli regulated by proportional–integral–derivative controller. This will allow us to further study the dynamics of growth of plant roots and changes in the ultrastructure of cells when the pressure and temperature factors are applied to the sample at the same time.
Optimization of iron dosage for microalgal biomass production as a feedstock for biofuel
Published in Biofuels, 2021
Probir Das, Mahmoud Ibrahim Thaher, Mohammed Abdul Quadir Mohd Abdul Hakim, Hareb Mohammed S.J. Al-Jabri, Ghamza Saed H.S. Alghasal
It was shown that iron starved Scenedesmus sp. doubled the carbon fixation into protein [28]. On the contrary, reduction in iron concentration was shown to reduce the soluble protein fraction in Cyclotella meneghiniana [29]. Furthermore, Menzyanova et al. (2009) showed contrasting dose dependent effects of iron on the protein content of Dunaliella viridis in four seasons [30]; even for the same iron concentration, protein content in D. viridis varied in different seasons. Similarly, mixed results were obtained in this study for both marine and freshwater microalgae.
In vivo imaging/detection of MRSA bacterial infections in mice using fluorescence labelled polymeric nanoparticles carrying vancomycin as the targeting agent
Published in Journal of Biomaterials Science, Polymer Edition, 2020
Araz Norouz Dizaji, Dan Ding, Tulin Kutsal, Mustafa Turk, Deling Kong, Erhan Piskin
The specific targeting properties of the CPDP@Van NPs against MRSA bacteria was investigated using Confocal Laser Scanning Microscopy (CLSM; Zeiss LSM 410, Jena, Germany). Quantitative analyses of the CLSM images were determined using Image Pro Plus software. The bacteria were centrifuged at 3000 rpm and the supernatant was removed. Then, 50 µl of glycerol were added and the bacteria for fixation purpose. Ten microliter of each sample was dropped on microscope slides images were taken with the confocal microscope upon excitation at 494 nm with the absorption of fluorescent signals around 531 nm.